Human pancreatic elastase I

Abstract

Human pancreatic elastase I can be obtained by genetic engineering.

We claim: 
     
       1. An isolated and purified DNA Sequence which encodes human pancreatic elastase I of the following amino acid sequence:   ______________________________________                                    
(N)--  Val Val Gly Gly Thr Glu Ala Gly Arg Asn Ser                        
Trp Pro Ser Gln Ile Ser Leu Gln Tyr Arg Ser Gly Gly                       
Ser Trp Tyr His Thr Cys Gly Gly Thr Leu Ile Arg Gln                       
Asn Trp Val Met Thr Ala Ala His Cys Val Asp Tyr Gln                       
Lys Thr Phe Arg Val Val Ala Gly Asp His Asn Leu Ser                       
Gln Asn Asp Gly Thr Glu Gln Tyr Val Ser Val Gln Lys                       
Ile Val Val His Pro Tyr Trp Asn Ser Asp Asn Val Ala                       
Ala Gly Tyr Asp Ile Ala Leu Leu Arg Leu Ala Gln Ser                       
Val Thr Leu Asn Ser Tyr Val Gln Leu Gly Val Leu Pro                       
Gln Glu Gly Ala Ile Leu Ala Asn Asn Ser Pro Cys Tyr                       
Ile Thr Gly Trp Gly Lys Thr Lys Thr Asn Gly Gln Leu                       
Ala Gln Thr Leu Gln Gln Ala Tyr Leu Pro Ser Val Asp                       
Tyr Ala Ile Cys Ser Ser Ser Ser Tyr Trp Gly Ser Thr                       
Val Lys Asn Thr Met Val Cys Ala Gly Gly Asp Gly Val                       
Arg Ser Gly Cys Gln Gly Asp Ser Gly Gly Pro Leu His                       
Cys Leu Val Asn Gly Lys Tyr Ser Leu His Gly Val Thr                       
Ser Phe Val Ser Ser Arg Gly Cys Asn Val Ser Arg Lys                       
Pro Thr Val Phe Thr Arg Val Ser Ala Tyr Ile Ser Trp                       
Ile Asn Asn Val Ile Ala Ser Asn --(C).                                    
______________________________________                                    
     wherein (N) represents (i) a Met residue,   (ii) Thr Gln Asp Leu Pro Glu Thr Asn Ala Arg or   (iii) any portion of the peptide of said (ii) with the proviso that the C-terminal Arg residue is present.   
     
     
       2. An isolated and purified DNA sequence which encodes a protein selected from the group consisting of proenzymes, and preproenzymes of the human pancreatic elastase of claim 1. 
     
     
       3. An isolated and purified sequence represented by the following formula: ##STR5## wherein X represents a stop codon (TAA, TGA or TAG); and the base sequence optionally has at its 5'-end ATG or a part or all of the sequence (5')-ACC CAG GAC CTT CCG GAA ACC AAT GCC CGC-(3'). 
     
     
       4. An isolated and purified DNA sequence which codes for a fusion protein comprising human pancreatic elastase I of the following amino acid sequence:   ______________________________________                                    
(N)--  Val Val Gly Gly Thr Glu Ala Gly Arg Asn Ser                        
Trp Pro Ser Gln Ile Ser Leu Gln Tyr Arg Ser Gly Gly                       
Ser Trp Tyr His Thr Cys Gly Gly Thr Leu Ile Arg Gln                       
Asn Trp Val Met Thr Ala Ala His Cys Val Asp Tyr Gln                       
Lys Thr Phe Arg Val Val Ala Gly Asp His Asn Leu Ser                       
Gln Asn Asp Gly Thr Glu Gln Tyr Val Ser Val Gln Lys                       
Ile Val Val His Pro Tyr Trp Asn Ser Asp Asn Val Ala                       
Ala Gly Tyr Asp Ile Ala Leu Leu Arg Leu Ala Gln Ser                       
Val Thr Leu Asn Ser Tyr Val Gln Leu Gly Val Leu Pro                       
Gln Glu Gly Ala Ile Leu Ala Asn Asn Ser Pro Cys Tyr                       
Ile Thr Gly Trp Gly Lys Thr Lys Thr Asn Gly Gln Leu                       
Ala Gln Thr Leu Gln Gln Ala Tyr Leu Pro Ser Val Asp                       
Tyr Ala Ile Cys Ser Ser Ser Ser Tyr Trp Gly Ser Thr                       
Val Lys Asn Thr Met Val Cys Ala Gly Gly Asp Gly Val                       
Arg Ser Gly Cys Gln Gly Asp Ser Gly Gly Pro Leu His                       
Cys Leu Val Asn Gly Lys Tyr Ser Leu His Gly Val Thr                       
Ser Phe Val Ser Ser Arg Gly Cys Asn Val Ser Arg Lys                       
Pro Thr Val Phe Thr Arg Val Ser Ala Tyr Ile Ser Trp                       
Ile Asn Asn Val Ile Ala Ser Asn --(C).                                    
______________________________________                                    
     wherein (N) represents (i) a Met residue,   (ii) Thr Gln Asp Leu Pro Glu Thr Asn Ala Arg or   (iii) any portion of the peptide of said (ii) with the proviso that the C-terminal Arg residue is present.   
     
     
       5. A host cell transformed with a vector comprising the DNA sequence of claim 1. 
     
     
       6. A host cell transformed with a vector comprising the DNA sequence of claim 3. 
     
     
       7. A host cell transformed with a vector comprising the DNA sequence of claim 4. 
     
     
       8. The host cell according to claim 5, wherein said vector further comprises expression regulatory elements operably linked to the human pancreatic elastase I encoded therein. 
     
     
       9. The host cell according to claim 6, wherein said vector further comprises expression regulatory elements operably linked to the human pancreatic elastase I encoded therein. 
     
     
       10. The host cell according to claim 7, wherein said vector further comprises expression regulatory elements operably linked to human pancreatic elastase I encoded therein. 
     
     
       11. The host according to claim 5, wherein the host cell is selected from the group consisting of E. coli, B. subtilis, yeast and a mammalian cell line. 
     
     
       12. The host cell according to claim 6, wherein the host cell is selected from the group consisting of E. coli, B. subtilis, yeast and a mammalian cell line. 
     
     
       13. The host cell according to claim 7, wherein the host cell is selected from the group consisting of E. coli, B. subtilis, yeast and a mammalian cell line. 
     
     
       14. A process for production of a human pancreatic elastase I, which comprises: (1) inserting the DNA sequence which encodes the human pancreatic elastase I of the following amino acid sequence:   ______________________________________                                    
(N)--  Val Val Gly Gly Thr Glu Ala Gly Arg Asn Ser                        
Trp Pro Ser Gln Ile Ser Leu Gln Tyr Arg Ser Gly Gly                       
Ser Trp Tyr His Thr Cys Gly Gly Thr Leu Ile Arg Gln                       
Asn Trp Val Met Thr Ala Ala His Cys Val Asp Tyr Gln                       
Lys Thr Phe Arg Val Val Ala Gly Asp His Asn Leu Ser                       
Gln Asn Asp Gly Thr Glu Gln Tyr Val Ser Val Gln Lys                       
Ile Val Val His Pro Tyr Trp Asn Ser Asp Asn Val Ala                       
Ala Gly Tyr Asp Ile Ala Leu Leu Arg Leu Ala Gln Ser                       
Val Thr Leu Asn Ser Tyr Val Gln Leu Gly Val Leu Pro                       
Gln Glu Gly Ala Ile Leu Ala Asn Asn Ser Pro Cys Tyr                       
Ile Thr Gly Trp Gly Lys Thr Lys Thr Asn Gly Gln Leu                       
Ala Gln Thr Leu Gln Gln Ala Tyr Leu Pro Ser Val Asp                       
Tyr Ala Ile Cys Ser Ser Ser Ser Tyr Trp Gly Ser Thr                       
Val Lys Asn Thr Met Val Cys Ala Gly Gly Asp Gly Val                       
Arg Ser Gly Cys Gln Gly Asp Ser Gly Gly Pro Leu His                       
Cys Leu Val Asn Gly Lys Tyr Ser Leu His Gly Val Thr                       
Ser Phe Val Ser Ser Arg Gly Cys Asn Val Ser Arg Lys                       
Pro Thr Val Phe Thr Arg Val Ser Ala Tyr Ile Ser Trp                       
Ile Asn Asn Val Ile Ala Ser Asn --(C).                                    
______________________________________                                    
        wherein (N) represents (i) a Met residue, (ii) Thr Gln Asp Leu Pro Glu Thr Asn Ala Arg or   (iii) any portion of the peptide of said (ii) with the proviso that the C-terminal Arg residue is present, into an expression vector;     (2) transforming a host cell with the expression vector;   (3) culturing the host cell under conditions which induce the expression of the DNA sequence; and   (4) isolating said human pancreatic elastase I from the culture medium.   
     
     
       15. The process according to claim 14, wherein the host cell is selected from the group consisting of E. coli, B. subtilis, yeast and a mammalian cell line.